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Human EGFR Antibody

R&D Systems, part of Bio-Techne | Catalog # AF231

R&D Systems, part of Bio-Techne
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AF231
AF231-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

CyTOF-ready, Dual RNAscope ISH-IHC Compatible, ELISA Capture (Matched Antibody Pair), Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Simple Western, Western Blot

Cited:

Co-Immunoprecipitation, ELISA Detection, ELISA Development, ELISA Microarray Development, IHC/IF, Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human EGFR
Leu25-Ser645
Accession # CAA25240

Specificity

Detects human EGFR in ELISAs and Western blots. In sandwich ELISAs, approximately 3% cross-reactivity with recombinant mouse EGFR is observed and less than 0.1% cross-reactivity with recombinant human (rh) ErbB2 and rhErbB3 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human EGFR Antibody

Detection of Human EGFR antibody by Western Blot.

Detection of Human EGFR by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and MDA-MB-231 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EGFR at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of EGFR antibody in A431 Human Cell Line antibody by Flow Cytometry.

Detection of EGFR in A431 Human Cell Line by Flow Cytometry.

A431 human epithelial carcinoma cell line was stained with Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.
EGFR antibody in A431 Human Cell Line by Immunocytochemistry (ICC).

EGFR in A431 Human Cell Line.

EGFR was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Human EGFR Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human skin

Flow Cytometry

0.25 µg/106 cells
Sample: A431 human epithelial carcinoma cell line

Immunocytochemistry

1-15 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed frozen sections of human skin

Immunoprecipitation

1 µg/mL
Sample: A431 human epithelial carcinoma cell line, see our available Western blot detection antibodies

Simple Western

12.5 µg/mL
Sample: A431 human epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and MDA‑MB‑231 human breast cancer cell line

Human EGFR Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human EGFR Biotinylated Antibody (Catalog # BAF231)
  • Standard: Recombinant Human EGFR Protein (Catalog # 1095-ER)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using AF231 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: EGFR

The epidermal growth factor receptor (EGFR) subfamily of receptor tyrosine kinases comprises four members: EGFR (also known as HER1, ErbB1 or ErbB), ErbB2 (Neu, HER2), ErbB3 (HER3), and ErbB4 (HER4). All family members are type I transmembrane glycoproteins that have an extracellular domain which contains two cysteine-rich domains separated by a spacer region that is involved in ligand binding, and a cytoplasmic domain which has a membrane-proximal tyrosine kinase domain and a C-terminal tail with multiple tyrosine autophosphorylation sites. The human EGFR gene encodes a 1210 amino acid (aa) residue precursor with a 24 aa putative signal peptide, a 621 aa extracellular domain, a 23 aa transmembrane domain, and a 542 aa cytoplasmic domain. EGFR has been shown to bind a subset of the EGF family ligands, including EGF, amphiregulin, TGF-alpha, betacellulin, epiregulin, heparin-binding EGF and neuregulin-2 alpha in the absence of a co-receptor. Ligand binding induces EGFR homodimerization as well as heterodimerization with ErbB2, resulting in kinase activation, tyrosine phosphorylation and cell signaling. EGFR can also be recruited to form heterodimers with the ligand-activated ErbB3 or ErbB4. EGFR signaling has been shown to regulate multiple biological functions including cell proliferation, differentiation, motility and apoptosis. In addition, EGFR signaling has also been shown to play a role in carcinogenesis (1 - 3).

References

  1. Daly, R.J. (1999) Growth Factors, 16:255.
  2. Schlessinger, J. (2000) Cell. 103:211.
  3. Maihle, N.J. et al. (2002) Cancer Treat. Res. 107:247.

Long Name

Epidermal Growth Factor Receptor

Alternate Names

EGF R, ErbB, ErbB1, HER-1

Entrez Gene IDs

1956 (Human); 13649 (Mouse); 24329 (Rat); 102138724 (Cynomolgus Monkey)

Gene Symbol

EGFR

UniProt

Additional EGFR Products

Product Documents for Human EGFR Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human EGFR Antibody

For research use only

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